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підручники. біохімія / Uchebnik_po_Biokhimii_E_S_Severin

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Ta6n1111.1111·3. lil3MeHeH111e cyMMapHoro 3apHAa aMMHOK111cnoT s 3as111c111MocT111 OT pH cpeAbl

CnJihHO KHCJiaH cpe,11,a

HeihpaJibHaH cpe,11,a

CnJihHO m.eno'luaHcpe,11,a

1. AMHHOKHCJIOTbl c ue,11,uccou,uupyrom.nMH pa,11,HI~aJiaMH

 

 

+H+

 

 

 

+oH-

 

NHa+ -CH-COOH

 

 

 

 

 

NHa+-CH-COO----- NH2CH - coo-

 

 

 

I

 

 

 

I

'

R

 

 

 

R

 

 

 

R

CyMMapub1ll. 3ap"1,II = +1

CyMMapub1ll. 3ap»,1:1 = 0

CyMMapnb1ll. 3ap»,1:1 = -1

2. AMHHOKHCJIOTbl c aHHOHHblMH rpynnaMH B pa,11,uKaJie

 

 

+H•

 

 

 

+OH-

 

NHa• -CH-COOH

 

 

 

 

NH3•-cH-COO-----

NH2-CH-COO-

 

 

 

I

 

 

 

I

'

CH2

 

 

 

CH2

CH2

I

 

 

 

I

I

COOH

 

 

 

coo-

coo-

CyMMapui,1ii 3ap»,1:1 = +1

CyMMapuhlii 3ap»,1:1 = -1

CyMMapnhlii 3ap»,1:1 = -2

3. AMHHOKHCJIOTbl c KaTHOHHblMH rpynnaMH B pa,11,uKaJie

 

 

+H•

 

 

 

+OH-

 

NH3•-cH-COOH

 

 

 

 

NH3+ - CH-coo----- NH2-CH-COO-

 

 

 

 

I

'

'

(CH2)4

 

 

 

(CH2)4

(CH2)4

I

 

 

 

I

I

NH3•

 

 

 

NHa•

NH2

CyMMapui,1ii 3ap»,II = +2

CyMMapni,1ll. 3ap}!,II = +1

CyMMapn1>1ll. 3ap»,1:1 = -1

,UHMhie )],JUI BbIIIOJIHeHHH HMH crreumpH'-IeCKHX <PYHKUHif. TaK, y-Kap60KcHrJIYfaMHHOBaH KHCJIOTa BXO)J,HT B COCTaB 6eJIKOB, yqaCTBYJOlll;IiX B CBepThIBamrn: KpOBH, H )],Be 6JIH3KO Jie)KaIUHe Kap6oKCHJihHbie rpyrrrrhI B HX CTpYJ<.T)'peHeo6xo,UHMhl ,UJIH

CBH3hIBattHH 6eJIKOBhIX cpaKTopoB c wottaMH CaH. Hapyrnettwe Kap6oKCHJTHPOBaHHH rJIYfaMaTa rrpwBOJJ,HT K CHH)KeHHIO CBepTbrnaeMOCTH KpOBH.

3HaqettHe rH,UpOKCHJlhHhIX rpyrrrr B COCTaBe Jlll3HHa ll rrpOJIHHa OITHCaHo B pa3,UeJie 15.

6. XUMu11ecKue pea«u,uu, ucnoAb3YeMble iJJlJl

o6napyJ1CeHUH a.MUHOKUCJlOnl

Crroco6ttocTh aMHHOKHCJIOT BCTyrraTh B Te HJTH HHbie XllMW-IeCKHe peaKUllll orrpe,UeJIHeTCH HaJIH- 'IHeMB HX cocTaBe cpyHKUHOHaJihHhIX rpyrrrr. TaK KaK BCe aMHHOKHCJIOThl, BXO,UHIUHe B COCTaB 6eJIKOB, CO,Uep)KaT y a-yrnepo,UHOro aTOMa aMHHOll Kap6oKCHJihHYJO rpyITilhl, OHH MOryT BCTYITaTb B xapaKTepttbie )],JU[ Bcex aMMHOKHCJIOT XHMWleCKHe peaKUHH. HaJIH'IHe KaKMX-Jitt6o cpyttKUHOHaJihHhIX rpyrrn B pa,UHKaJiax llH)l,HBMJJ,YaJihHhlX aMHHOKHCJIOT orrpe,UeJIHeT 11x crroco6HocTh BCTYrraTh B crreuttcpW-IHbie ,UJIH ,UaHHhIX aMHHOKHCJIOT peaKUMM.

HHHfll.UPHHOWUI peaKIUHI ua a-aMHHOKHCJIOThI

)l.JIH o6ttapy)KeHHH H KOJIH'IeCTBeHHOfOorrpeJJ,eJieHMH aMMHOKMCJIOT'HaXOJJ,HIUMXCH B paCTBO-

pe, MO)KHO HCITOJlh30BaTh HHHfH,UpHHOBYIO peaKUHIO.

3Ta peaKUHH OCHOBaHa Ha TOM, 'ITO 6ecuBeTHhIH HHHfH,!1,pHH, pear11pyH c aMHHOKHCJIOTOH, KOH)l,eHCHpyeTCH B BH,Ue JJ,HMepa qepe3 aTOM aJOTa, OTIUeITJIHeMbIH OT a-aMirnorpyrrrrhI aMHHOKHCJIOTbl. B pe3yJihTaTe 06pa3yeTCH rrttrMeHT KpacHocp110JieT0Boro UBeTa. 0,UHOBpeMeHHO rrpOMCXO)l,MT JJ,eKap60KCHJIMPOBaHMe aMHHOKHCJIOThI, 'ITOrrpllBOJJ,MT K o6pa30BaHHIO C0 2 l1 COOTBeTCTBYJOIUefO aJih,Uer11,Ua. HttttrH,UpHHOBYIO peaKUHIO wttpoKo HCITOJih3YJOT rrp11 113yqeH1111 rrepBw-rnott cTpYJ<.TYphI 6eJIKOB (CM. CXeMY HH)Ke).

TaK KaK HHTeHCHBHOCTh oKpacKH rrporropu110HaJihHa KOJIH'IeCTBYaMMHOKHCJIOT B pacrnope, ee HCilOJib3YIOT )lJIH H3MepeHMH KOHUeHTpaUMM a- aMHHOKHCJIOT.

2 x

 

 

 

+

 

 

 

 

 

 

 

 

 

0

 

0

 

II

 

 

II

 

(irc,C=N-C ,,.c--f)

+

V-c/

I

'C-V

 

II

H

 

II

 

0

 

0

 

HMHrMAPMHOBaH peaKLIMH, MCnOnb3yeMaH AJIH onpeAeneHMH a-aMMHOKMCOOT

14

CneIUt<l>ffllecKHe peaKIUtH

ua OT)J;MbHbie aMHHOKHCJIOTbl

KaqecnieHHoe 11 KOJI11qecTBeHHoe orrpe)J,eJieH11e OT)J,eJibHbIX aMWHOKHCJIOT B03MO)KH0 6Jiaro)J,apH HaJI11qmo B 11X pa)J,11KaJiax oco6eHHbIX ¢>YHKI.J,110HaJibHbIX rpynn.

Apr11H11H onpe)J,eJIHIOT c rroMOIUbIO KaqecrneHHoif peaKI.J,11.11 Ha ryaHM)J;MHOBYJO rpynrry (peaKUWH CaKaiym), a uwcTettH BhIBBJUIIOT peaKUtteti <l>oJIH, crreumpttqHOti Ha SH-rpynny )J,aHHOM aMMHOKHCJIOTbI. Hanwrne apoMaTMqecKMx aMttHOKMCJIOT B pacrnope orrpe)J,eJIHIOT KcamorrpoTe11HOBOif peaKuweti (peaKI.J,MH HMTpoBaHHH), a Han11q11e rlf)J;poKCHJibHOi1 rpynIIbI B apoMaTttqeCKOM KOJibU:e TMp0311Ha - c noMOIUbIO peaKu1111 MHJIJioHa.

Ji. IIEIITlf,IJ;HAH CBH3h. CTPOEHHE

II BHOJIOfHqECKHE CBOHCTBA IIEIITH,I(OB

a-AM11HOK11CJIOTbI MOryT KOBaJieHTHO CBH3bIBaTbCH )J,pyr c )J,pyroM c IIOMOIUblO rreIITM)J;HbIX CBH3eti. IleIITM)J;HaH CBH3b o6paJyeTCH Me)K,11,y o:-Kap- 60KC11JibHOM rpyrrnoti 0)1,HOM aMttHOKHCJIOTbl M a-aM11HOrpynrroti )J,pyroti, T.e. HBJIHeTCH aMM)J;HOM CBH3bl0. Ilptt 3TOM IIP011CXO)J,11T OTIUeIIJieHtte MOJieKYJibl BO)J,bl (CM. cxeMY A).

1. Cmpoenue nenmuiJa

KoJitt'Iecrno aMMHOKHCJIOT B cocrnBe rreIITM)J;oB MO)KeT CHJibHO BapbttpOBaTb. IleIITM)J;bI, CO)J,ep%aIUMe )J,O 10 aMWHOKMCJIOT, Ha3bIBa10T OJilll'ODeIIl'H)J.bIqac. -

TO B Ha3BaHMH TaKWX MOJieeyJI yKa3bIBalOT KOJilfqeCTBO BXOMIUMX B COCTaB OJittrOIIeIITM)J;a aMHHOKHCJIOT: Tp1menTM)J;, rremarremlf)J;, oKTarrerrTM)J; tt T.)J,.

IleIITM)J;bI, coaep%aIUMe 6oJiee 10 aM11HOKMCJIOT Ha3bIBaIOT «DOJIHDellfU)U>I»' a IIOJrnITeIITM)J;bI, COCTOH~ mtte tt3 6oJiee qeM 50 aMHHOKHCJIOTHbIX ocrnTKoB 06bJqH0 Ha3b1Ba10T 6eJIKaM:tt. 0)1,HaKO 3TII Ha3BaH~

YCJIOBHbI, TaK KaK B JIMTepaType repMttH «6eJIOK» qaCTO ynoTpe6JIHIOT )J,JIH o603Ha'IeHHHIIOJI11IIeIITM)J;a, CO)J,ep%a!Uero Mettee 50 aMHHOKHCJIOTHbIX OCTaTKOB. HanpttMep, ropMoH rJIIOKaroH, coCTOHIUttti tt3 29 aMMHOKHCJIOT'Ha3bIBaIOT 6eJIKOBbIM ropMOHOM.

MoHoMepbI aMHHOKWCJIOT, BX0)1,SIJ.Ul1X B cOCTaB 6eJIKOB, Ha3bIBaIOT <~aMHHOKHCJIOillbie ocraTKH». AMMHo-

KMCJIOTHbili OCTaTOK, HMeIOIUHti CBOOO)J,HylO aMMHO-

rpynny, Ha3bIBaeTCH N-KOHUeBhIM 11 Ill1IIIeTCH cJieBa a HMelOIUMH CB060)lHYJO a-Kap60KCHJihHyIO rpyn~ rry - C-KOHUeBblM 11 IIMIIIeTCH crrpaBa. neIITM)J;hI IIMlllYfCH tt IJ1ITa10TcH c N-KOHUa. UeIIh rromopHIOIUMXCH aTOMOB B rroJittrreIITM)J;HOM u,errtt -NH-CH-CO- HOCITT Ha3BaHMe «ne~m ocroe» (cM. cxeMy E).

Ilp11 Ha3BaHMH IIOJIHrremlf)J;a K COKpameHHOMY Ha3BaHMIO aMHHOKHCJIOTHbIX OCTaTKOB ao6aBJUIIOT cy<P<PttKc -HJI, 3a HCKJI10qeH11eM C-KOHU:eBoif aMwHOKMCJIOTbI. HarrpttMep, TeTparrerrTtt)J, Cep-fJitt- Ilpo-AJia 'IMTaeTCHKaK cepMJirJiilllMJIIIPOJIMJiaJiaHMH.

IleIITM)J;HaH CBH3h, o6paJyeMaH 11MHHOrpynrroti

npOJIMHa, OTJ1WiaeTCH OT llPYfHX rremlf)J;HbIX CBH3eti, TaK KaK aTOM a30Ta rreIITM)J;HOM rpyrrrrhl CBH3aH He c BO)J,OpO)J,OM, a c pa)J,11KaJIOM.

IleIITM)J;bI pa3JIHlla10TCH no aMHHOKHCJIOTHOMY coCTaBY, KOJIH'!eCTBYM IIOPHllKY COe)lMHeHHH aMMHO-

+

 

r-----,

 

H3N-CH-CO-NH-CHTC0-N +CH-CO-NH-CH-C00-

1

I

~---.,- 1 I

I

CH2

H

H2C CH2

CH3

I

 

'- /

 

OH

 

CH2

 

Cep1mrnM1.1MnnponMnanaHMH

H~-CH-CO-NH-CH-COOH

 

 

 

I

'---,.-------' I

 

 

 

R1

l

R2

 

06pa3oBaHa-te .1:111nenrn.i:1a

 

nenT11.QHaR

 

CxeMa A. 06pasoeaHMe AMnenTMA8

 

 

 

CBR3b

 

 

 

 

 

 

Pat:1111Kanb1 aMHHOK111cnoT (6oKoBaR 1..1enb}

 

 

~

l

~ ---- .

 

 

 

R2

R3

 

 

 

 

I

I

I

 

 

I +H3N CH CO-NH-CH-CO-NH-CH-CO ....

NH-CH- C00-1- nenrn.QHbt'1

T

 

' --- v ----- '

 

T

OCTOB

 

 

 

 

AMHHOKHCnOTHbl'1OCTaTOK

 

 

N-KOHBl..l

 

 

C-KOHBL\

 

 

 

 

 

CxeMa 6. CTpOeHMe nemMAOB

15

1-1).

KHCJTOT. Cep-fHc-npo-A.rra 11 A.rra-Tipo-r11c-Cep -

.[(Ba pa3HhIX nemruia, HeCMOTp5I Ha TO, 'ITOOHH HMeIOT O)lHHaKOBhie KOJIWieCTBeHHhIH 11 KaqecTBeHHhlH coCTaBhl aMHHOKHCJTOT.

2. XapaKmepucmuKa nenmuiJuoii C8H3U

nerrTH)lHa5I CB5I3h HMeeT xapaKTepHCTHKY qac-

TH'lHO)lBOHHOH CBJl311, n03TOMy Otta KOpoqe, qeM OCTaJThHhie CB5I3M nenTH)lHOfO OCTOBa, 11 BCJTe)l- CTBHe 3TOro MaJTO ITO,LJ,Bl-DKHa. 3JTeKTPOHHOe CTpoeHHe nenTH)lHOH CB5I311 orrpe)lemreT ITJTOCKYIO )!(e- CTKYIO CTPYKTYPY rrem11,LJ,HOH rpynrrhI. TIJTOCKOCTH rrerrrn,LJ,HhIX rpynrr pacnoJTO)J(eHhI no,LJ, yrJToM ,D,pyr K ,D,pyry (p11C.

CBJ13b Me)[()ly a-ymepo)lHbIM aToMOM 11 a-aM11- ttorpynrrot'!11JT11 a-Kap6oKC11JTbHOH rpynnoti cnoco6Ha K CB060llHbIM Bpaw:emrn:M (xorn orpatt11Y.e-

Ha pa3MepoM 11 xapaKTepoM pa,LJ,MKaJTOB), '-I.TO no3Bom1eT nommenTH1lHOH uem1 np11tt11MaTb pa3JT11qtth1e Kottqmrypau1111.

TienTH,LJ,Hbie CB5I311 o6bIY.HO pacnOJTO)!(eHbl B TpaHC-KOHCf)l1rypaUH11, T.e. a-yrnepO,D,Hbie aTOMbl pacnoJTaraIOTCH no pa3Hbre CTOpOHhI OT nenTH)l- HOM CBH311. B pe3yJTbTaTe 6oKOBhie pa,LI,ttKaJThI aMHHOKHCJTOT ttaxo,LJ,HTCH tta tta1160JTee y,LJ,aJTeHHOM pacCTOHH1111 ,D,pyr OT )lpyra B npOCTpaHcTBe (p11c. 1-2).

CTenTH,LJ,Hble CB5I311 oqeHh rrpoqHhl 11 CaMOTip0113-

BOJlhHO He pa3phmaIOTCH rrp11 HOPMaJTbHbIX ycJTOBHHX, cyw:eCTBYIOW:HX B KJTeTKax (HeMTPaJThHaH cpe,LJ,a, reMneparypa TeJTa). B JTa6oparopttbIX ycJTOBMHX rl1)l- poJTH3 neITTH,LJ,HblX CB5I3et) 6eJTKOB npOBO)l5IT B 3arraHHHOH aMrryJTe c KOHUeHTpl1pOBaHHOH (6 MOJTh/JT) COJT5IHOH KHCJTOTOH, np11 TeMrreparype 6oJTee 105 °C, np11Y.eM TIOJTHbIH rn)lp0JT113 6eJTKa ,LJ,O CB060,LJ,HbIX aM11HOKHCJTOT rrpoxo,LJ,m np11Meptto 3a cyrKH.

B )J(J1BbIX oprattl13Max nerrT11,LJ,Hb1e CB5I311 B 6eJT-

Kax pa3pbIBaIOTC5I c TIOMOWbIO cneuMaJThHblX rrpoTeom1rnY.eCKl1X cjlepMeHTOB (oT attrn. protein - 6e-

JTOK, lysis - pa3pywett11e), Ha3bIBaeMhIX TaK)J(e rrpoTea3aJ1.rn, 11JT11 rrenTl1)lr11,LJ,pOJTa3aM11.

Ll,JT5I 06Hapy)J(eH115! B pacrnope 6eJTKOB 11 nenT11llOB, a TaK)J(e .wrn 11X KOJT11qeCTBeHHOfO onpe,D,eJTemrn 11CTI0Jlb3YIOT 611ypeTOBYIO peaKU11!0 (nOJTO)!(H- TeJTbHblH pe3yJTbTaT llJ15I Bew:ecTB, CO,D,ep)J(aw:11x B CBOeM COCTaBe He Mettee llByx neTITH,Ll,HbIX CBH3ei1).

3. Euo.n.ozurtecKaa po.n.b nenmuiJoe

B opraHH3Me Y.eJToBeKa Bbipa6aThIBaeTC5I MHO- )J(ecrno nenTH,LI,OB, yY.aCTBYIOW:l1X B peryJTHUJ.111 pa3Jll1Y.Hb!X 6ttOJTOrttY.eCKl1X npoueccOB 11 o6JTa,LI,aIOW:HX BhICOKOH cPVI3110JTOrnqecKOH aKT11BHOCTblO.

 

®

 

a-YrnepO.QHbliil

 

 

 

 

 

 

H j

a TOM

®

 

 

 

 

0

 

I

 

0

I

 

~

 

 

 

 

 

 

 

II

 

 

I

II

 

 

 

c

 

CH

c

/N"-...

/c,, /

CH

"c

/N"-...

 

 

/ "N /

 

 

CH

N

 

 

 

 

I

 

 

II

 

I

 

II

 

 

 

H

 

 

0

 

H

 

0

 

 

 

 

 

 

 

®

 

 

 

r

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

,/'

 

 

nnoCKOCTb

 

 

 

Pap,11Kan

 

 

nem11p,Hoiil

 

 

aMl1HOK11CilOTbl

 

 

 

 

 

 

rpynnbl

 

 

 

 

 

 

 

P1o1c. 1-1. nnocKOCTM pacnono)l(eHMH nenTMAHblX rpynn M a-yrnepOAHblX aTOMOB B npOCTpaHCTBe.

H

0

®

H

0

I

II

I ,,H

I

11

,.. N""- /

C ""- /

C""- /

N""- /

C ""-

C

N

C

C

 

H/ I

I

II

H/1

 

®

H

0

®

 

PMC. 1-2. TpaHC-KOH!j>Mrypa1.1MH nenTMAHblX CBH3eiii. <PyHKl..lli1· OH3flbHbl6 rpynnbl -CO- 11 -NH-, o6pai3ytol.L.!1'16nenTll1AHbl6 CB5131'1, He li10Hl'1311pOBaHbl,HO non51pHbl, 111 MOryT Y43CTBOBaTb B o6pa30BaHl'1111BOAOPOAHblX CB5136~.

KoJT11Y.eCTBO aMttttoK11CJTOTHbIX ocTaTKOB B CTPYKType 6110JTor11qecK11 aKT11BHbIX nenT111lOB MO)J(eT Baph11poBaTh OT 3 1lO 50. K O,LJ,HHM 113 caMbrx «MaJTeHbK11X» rrerrT11,LJ,OB MO)J(HO OTHeCnI Tl1-

peOTporr11H-pl1JTl1311Hf-rOpMOH 11 rnyraTHOH (Tpl1nenT11,Ll,bl), a TaK)J(e 3HKecjla;111HbI, 11Me10w:11e B cBoeM cocrnBe 5 aM11ttoK11cJToT. OllttaKo 6oJTbW11Hcrno 6110JTOfHY.eCKl1 aKT11BHbIX nenTH,LJ,OB 11MeeT B CBOeM COCTaBe 6oJTee 10 aM11HOK11CJTOT' Hanp11Mep

ttet'!porrem11,LJ, Y (peryJT5ITOp arrneTttTa)

C01lep)J(l1T

36 aM11ttoKHCJTOT, a KOpT11Ko1rn6ep11H -

41 aM11-

HOK11cJT0Ty.

HeKOTopb1e 113 nenT111lOB, B Y.aCTHOCTH 6oJTbWttHCTBO nenT11,Ll,Hb!X ropMOHOB, COllep)J(aT nenTH.llHhie CB5I311, o6pa3oBaHHbie a-aMHHOrpynrroii 11 a-Kap60KC11JlbHOH rpynnot'!COCellH11X aM11HOK11C-. JTOT. KaK npaB11JTo, ott11 c11ttTe311pyIOTC5I 113 tteaKTMBHhIX 6eJTKOBhIX npe,D,wecTBeHHHKOB, B KOTOpblX cneu11¢>11Y.ecK11e npoTeoJT11T11qecK11e cjlepMeHTW pa3pywaIOT orrpe,D,eJTeHHhie nenTttJJ,Hhre CBH311.

AHrnoTeH311H lI - 0Krnncml1)l, o6pa3y10w:11t'!- c51 113 Kpynttoro 6CJTKa nJTa3Mbl KPOB11 aHrHOTeH-

16

13).

3HHOreHa B pe3yJihTaTe ITOCJie!IOBaTeJibHOfO 11,eMCTBIDI 11,Byx ITPOTeOm1TH'IeCKHX<PepMeHTOB.

IlepBbIM ITpOTeomrrw1ecKHM <PepMeHT pemrn oTmeITmreT OT aHr110TeH31-rnoreHa c N-KoHua ITeITTHlJ,, C011,ep)l(amntt 10 aM11HOKHCJIOT, Ha3bIBaeMblM aHr110TeH311HOM I. BTopott ITpOTeoJinn1qecK11M <PepMeHT Kap60KCH11,HITeITTH11,ttnITeITTJi,l::(a3a OTmeITJHieT OT C-KoHua attrnoTeH3nHa I 2 aMnHoKHcJioThI, B pe3yJibTaTe qero 06pa3yeTC5! 6110JIOrn:'-1eCKl1 aKTl1BHhIM attr110TeH311H II, yqacrnyrom11if B peryJI51I.U1H A)], 11 BOlJ,HO-COJieBoro o6MeHa B opramnMe (cM. cxeMy A).

01lHaKO B HeKOTOpbIX 6HOJIOrtt'leCKHaKTHBHbIX ITeITT11aax MOryT coaep)l(aThC51 Jin6o tteo6hI'-IHhie aMl1HOKHCJIOThl, JIH60 cymeCTBOBaTh He06bI'-IHbie CB513H Me)l()l;y aMHHOKHCJIOTaMl1, He BCTpe'-1aIOlllH- ec51 B 6eJIKax.

TlpiIMep ITeITTnaa, coaepxrnmero Heo6bI'-IHYIO 11,JI51 6eJIKOB CB513h Me.)K)J;y aMHHOKMCJIOTaMl1, - TPHITeITTHll rnyrnTHOH, ITOCTpoeHHhIH 113 rnyrnMaTa, UHCTel1Ha 11 rJIHUHHa (CM. cxeMy :0).

N-KOHUeBaH aM11HOKl1CJI0Ta rJIYTaMaT cmnaHa co BTOpoif aMHHOKHCJIOTOM u11cTeHHOM He qepe3 a-Kap6oKCttnhHYJO rpyITny, a '-lepe3y-Kap6oKcHJih- HYIO rpyrrITy ero paa11KaJia. fJIYTaT110H - III11p0Ko

pacITpocTpatteHHhIM ITem11D: opratttt3Ma lfeJioBeKa. Ott MO)J(eT 6bITb 11cITOJih30BaH B OKHCJIHTeJibHOBOCCTaHOBHTeJihHbIX peaKLI,1151X KaK 11,0HOp H aKuenTOP 8011,0PO!Ia H He06X011,HM !IJIH pa60Tbl p5111,a <PepMeHTOB.

<l>yHKUHH ITeITTHlJ,OB 3aBHC51T OT HX ITepBH'-IHOM CTPYKTYPhI. AttrnoTeH3HH I ITO cTpyKType Olfettb ITOXO)l( tta aHrnoTeH3HH 11 (ttMeeT TOJihKO !IBe ao-

ITOJIHHTeJihHhie aMHHOKl1CJIOThI c C-Kottua), HO ITPH 3TOM He 06Jia11,aeT 6ttOJIOfl1'-leCKOHaKTHBHOCTblO.

J13MeHeH11e B aMHHOKHCJIOTHOM COCTaBe ITeITTl111,0B 'laCTOnp11B011,HT K ITOTepe O!IHl1X 11 B03Hl1KHOBeHHIO !IPYfHX 6110Jior11'-leCKHX CBoil:cTB. B Kalfecrne npnMepa MO)J(HO paccMOTpeTh CTPYKTYPY 11 CBOMCTBa 11,BYX ITeITTHlJ,HbIX ropMOHOB - OKC11TOll,11Ha n Ba3oITpeccntta.

B rHITOTaJiaMyce OKCHTOUHH 11 Ba30ITpecCHH o6pa3YJOTC51 B pe3yJihTaTe lfaCTl1'-IHOfO (orpaHl1'-leH- HOro) npoTeomna 6oJiee KPYITHbIX 6eJIKOBbIX ITpe!I- wecTBeHH11KOB. 113 fHITOTaJiaMyca ITO HepBHblM BOJIOKHaM 3TH ropMOHhI BHYTPH ceKpeToptthIX rpaHYJI rrepeMemaIOTC51 B HepBHbie OKOH'lamrnaKCOHOB, Haxo11,51mnxc51 B 3al1,Heif !IOJie rnrro<P1na. IlocJie aeifcTBH51 crreu11<P11YecKHx CTHMYJIOB 3TH ropMoHhI BhI11,emr10TC5! B KpoBb (cM. cxeMy A Ha c.

2

3

4

 

5

6

7

8

9

10

11

 

N-KOHel..I Acn -Apr- Ban-T111p-V1ne- r111c - npo- <lleHr111c -ne~-Ban-nenrn,[I C-KOHe1..1

 

 

 

 

AHn10T0H3111HoreH

 

 

 

 

 

 

 

 

 

 

 

 

1 PeHlllH

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

2

 

3

4

5

6

 

7

8

9

10

 

 

Acn -Apr - Ban - T111p-V1ne- r111c - npo- <lleHr111c - ne~

+ nemlll,[I

 

 

 

 

 

AHrl·10T0H3111H I

 

T

 

 

 

 

 

 

 

 

 

 

1

Kap60Kc111,Q111nenT111,Q111nnenT111.Qa3a

2

 

3

4

5

6

 

7

8

 

 

 

 

 

Acn -Apr - Ban - T111p-V111e- r111c - npo - <lleH

+

 

r111c -ne~

CxeMa A

 

 

 

AHr1110TeH3111H

II

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

rny

 

 

 

 

l..l111c

 

 

 

r11111

 

 

 

 

 

 

 

 

~~

 

 

 

 

 

 

 

..H3N-CH- CH-CH2-CHrCO-NH-CH -CO-NH-CH2-C00-

1

 

 

 

 

 

 

 

I

 

 

 

 

 

 

COO

 

 

 

 

 

 

 

CH2

 

 

 

 

 

 

 

 

 

 

 

 

 

 

I

 

 

 

 

 

SH

CxeMa Ei. Tp1menTH~ rnYTaTHOH ("trnyraMHm~HCTeHHHnrnMLl.HH)

17

OKcMrn11:irn M Ba3orrpeccMH B cBoetl CTPYKTYPe MMeIOT MHOro o6mero:

o6a co.uep)l(aT 9 aMMHOKHCJIOTHbIX OCTaTKOB;

7 aMMHOKHCJIOTHbIX OCTaTKOB M3 9 H,UeHTWIHbI;

2 OCTaTKa UHCTeHHa coe,UHHeHbl ,llMCYJibcPH.llHOH CB5I3bIO;

•Ha C-KoHue rrerrTH,UOB a-Kap6oKCHJibHa51 rpyrrrra r.rryTaMaTa aMH,UMpoBaHa.

HecMOTp51 Ha tte6onb11rne OTJIWIH51 B rrocne.uoBaTeJibHOCTM aMMHOKHCJIOT ( 3aMeHbl aMMHOKHCJIOT B ITOJIO)[(eHJUIX 3 M 8) 3TM ropMOHbl CHJibHO OTJIM- 'IaIOTC51 no cPH3MOJIOrMtiecKoMy .uertcrnmo. TaK, OKCMTOUMH BbI,UeJI51eTC51 B KpOBb BO BpeM51 KOPMJieHH51 pe6eHKa, Bbl3bIBaeT COKpameHHe MH03IIMTeJIMaJibHbIX KJieTOK rrpOTOKOB MOJIO'IHbIX)KeJie3 M CTMMYJittpyeT BbI,UeJieHMe MOJIOKa. KpoMe Toro, OKCMTOUHH BJltt51eT Ha fJia,Il,KYIO MYCKyJiaTYPY MaTKH BO BpeM51 pO,llOB, Bbl3bIBa51 ee COKpameHMe.

B OTJIH'IMe OT OKCMTOUMHa, OCHOBHOe cPH3HOJIOrH'IeCKOe .uetlcTBMe Ba3onpeccwHa - YBenw-1e- HHe pea6cop6um1 BO,Uhr B rrotIKax rrpM yMeHbIIIeHMM AU MJIM o6oeMa KPOBM (no3TOMY .upyroe Ha3BaHMe 3Toro ropMOHa - aHTM,UttypeTtttieCKHH). KpoMe Toro, Ba3orrpeccHH BbI3hIBaeT cy)l(eHtte fMK cocy,UOB.

HHTepeCHO OTMeTHTb, 'ITO HaJIH'rneB TTOJIO)[(e- HHM 8 OCHOBHOH aMllliOKHCJIOTbI Ba)KHO ,11/151 rrp051BJieHJUI alfl'lUUfYPeTH'IeCKOMaKTHBHOCTM, a aMMHOKHCJIOTbI c rn.upo¢o6HbIM pa,UMKaJIOM B ITOJTO)[(eHMH

3 - ,1lJT5I COKPaIIIeHH51 fMK.

TaK KaK nerrTH,UbI - MOll.lHhie peryAATOPbI 6HonorMtiecKMX rrpoueccoB, Mx MO)[(HO 11crroJib30BaTh KaK neKapcrneHHbie rrperrapaTbir. OcHOBHoe rrperr5ITcrn11e )l,JUI TepaneBTH'IeCKOroMCIIOJib30BaHH51 - HX 6bICTpOe pa3pyIIIeHMe B opraHH3Me. 0,UHMM M3 Ba)[(- HeHIIIMX pe3yJTbTaTOB HCCJTe,UOBaHHM 51BJI51eTC51 He TOJibKO H3yqeHHe CTpYJ<TYpbl rrerrTH,llOB, HO M rronytieHHe CHHTeTH'!eCKMX aHaJIOfOB rrpHpO,llHbIX rrerrTH,UOB c ueneHarrpaBJieHHbIMH H3MeHeHH51MH B HX CTpYKType H cPYHKUM51X.

HarrpttMep, c11Hre311poBaH rrerrrH,U l-.ueJaMMH0- 8-D-apmH11H-Ba3orrpecc1rn (Ll,AB), CTpYJ<TYPa KOToporo rrpe.ucraBJieHa Ha cxeMe E.

B CTPYKTYPe 3Toro rrerrTH,Ua (rro cpaBHeHMIO c BaJorrpecc11HoM) HeT aMttHorpyrrrrhI Ha N-KoHue, H BMeCTO L-aprMHHHa B Il()JlO)l(eHMM 8 CTOMT D-aprnHHH. TaKoti CMHTeTH'ICCKtttirrerrTH.ll o6na-

.uaer TOJTbKO aHTH,llttypeTH'IeCKOH aKTHBHOCTbIO H XHMH'IeCKHYCTOH'-IMB, T.e. rrpH BBe)JeHHH B opraHH3M BbI3hrnaeT )JJTMTeJihHYIO peaKUHIO. TaKoil HccyccrneHHbIM aHanor ropMOHa (no cpaBHeHHIO c rrpMpO)JHbIM) 6onee 3¢¢eKTHBCH rrptt Jie'IeHMMropMOHaJibHOM He)JOCTaTO'-IHOCTH.

OrKpbITbie tt tt3yqeHHbie B HacTomuee BpeM51 rrerrTH,llbI MO)[(HO pa3,UeJIHTb Ha rpyrrnbl ITO HX ocHOBHOMY cPM3HOJIOrtt'IeCKOMY)JeHCTBttIO:

rrerrTM)JbI, o6Jia,IJ,aJOIIWe ropMOHaJibHOM aKTHBHOCTblO ( OKCH:TOUMH, Ba3onpecCHH, PMJIM3HHrropMOHbl rnnoranaMyca, MeJiaHOUHTCTMMYJIHPYIOll.lHH ropMOH, rnIOKaroH tt JJp.);

 

2

3

4

5

6

7

8

9

 

N-KOH91..1

NHzL1111c- T111p-V1ne- lnH-Acn- L.1111cnpo- ne~ - lnyCONH2

C-KOH91..1

 

L __ s - s ----

 

 

 

 

 

 

 

OKc111To1..1111H

 

 

 

 

 

 

2

3

4

5

6

7

8

9

 

N-KOH91..1

NH2- L.1111c- T111p-Cl>eH- lnH-Acn- L1111c- npo-Apr - lnyCONH2

C-KOH91..1

 

L__s-s---~

 

 

 

 

Ba3onpecc111H

CxeMa A

2

3

4

5

6

7

8

9

N-KOH91..1 H-L.1111c-T111p-Cl>eH-lnH-Acn-L.1111c-npo-Apr - lny-CONH2 C-KOHel..I

L _ s -- s

o

8-D-apr111H111H-sasonpecc111H

CxeMa Ei

18

1-3).

nenTHJJ:hI, perym1py10m11e npouecch1 mm.1,eBapeH115l (racTpHH, XOJieUHCTOKHHl1H,. Ba3011HTeCTl1HaJibHblll rrerrnm:, )l(eJTY,!IO'-IHbJH:l1Hrn611py- JOIUl1H rremHJJ: 11 ,np.);

nenTHJJ:hI, peryn11pYJOW,11e Tottyc cocy,noB 11 A,LJ.

(6pan11KHH11H, Kari11,n11H, attrnoTeH311H II);

• nenTH.LI,hI, pery1rnpy10m11e anneT11T (nenT11H, Hei1porrenTl1).l, Y, MeJiaHou11TcT11Myn11py10m11t1 ropMOH, ~-3H).l,OpqHrnhI);

nenTHJJ:hI, 06na,n;a10w,11e 06e36on11Ba10m11M ).l,ettCTB11eM (3HKe<l:JaJil1Hbl 11 3H,Uop¢11Hbl 11 ).l,pyrne OITl10HJJ:Hhle rreITTHJJ:bl). 06e360Jil1BaIOUJ,11H 3W- <l:JeKT 3THX rreITTHJJ:OB B COTHl1 pa3 rrpeBOCXO).l,HT attarihre311py10IUH:H 3<l:J¢e1<T Mop¢1rna;

rrerrTHJJ:hI, yqacrny10m11e B perymn.1,1111 BhIClllett HepBHOH ,Ue51TeJibHOCTl1, B 6110Xl1MH'IeCKl1X rrpoueccax, cmI3aHHhIX c MexaHl13MaMH ctta, o6y4eHl15l, rraM51Tl1, B03Hl1KHOBeHl151 llYBCTBa

cTpaxa 11 T .).],.

0,nHaKO TaKOe ).l,eJieH11e rreIITl1).l,OB Kpati.He ycJIOBHO. IloHBl1Jil1Cb ).l,aHHhie 0 TOM, 4TO MHOrHe neIIT11).l,bl o6Jia,UaIOT IIIHPOKl1M cneKTpOM ,neti.- CTBl151, TaK, MeJiattou11TCT11Myn11py10m11tf ropMOH, IIOM11MO CT11MYJI51U1111 mtrMeHTOo6pa30BaHl15l, yqacrnyeT B peryn51u1111 anneTl1Ta (BMecTe c nenTl1HOM no,naBJI5leT IlOTpe6JieH11e TI111Ul1 11 51BJI5leTC51 attrnrott11CTOM tteti.porrerrT11Aa Y). B TO )Ke BpeMH ~-3H)J,OPW11Hhl, KPOMe attaJibre3Hpy10mero 3<l:J<l:JeKTa, - c11Hepr11cTbI HetfponenT11na Y, T.e. yc11JI11BaIOT n0Tpe6nett11e rr11w,11. Orr11caHHb1ti.

Bblllle Ba3onpecCHH, KPOMe aHTHJIIIypeT11'-!eCKoro

11 cocy).l,oc~11BaIOmero netfcTBl1H, ttMeeT cBoii-

CTBO YJIY'-!lllaTbrraMHTb.

D 7 7

 

7 7

7

~-N-C-C-N-C-C-N-

1

11

I

II

H

0

CH30

II. CTPYKTYPA 6E11KOB

IlenTHJJ:Hhie uerrH CO)l,ep)KaT ).l,eCHTKl1, COTHH 11 TbICWIH aMHHOKHCJIOTHbIX OCTaTKOB, coe).l,HHeHHbIX rrp04HbIMl1 rrerrTH)],HblMl1 CB51351Ml1. 3a C'-leT BHYTPl1MOJieJ<YAAPHhIX B3attMOAettcTBHi1 6eJIKH o6pa3y-

JOT orrpe,IIeJieHHYIO npocTpaHCTBeHHYIO CTpyrcrypy, Ha3hIBaeMyJO «KOHCl>opMau;m1 6eJIKOB». JlHHei1Ha51 ITOCJie)lOBaTeJibHOCTb aMHHOKHCJIOT B 6eJIKe co,nep-

)1(11T HH<l:JopMaumo 0 ITOCTpOeHl1H TpexMepHOH rrpoCTpattcTBeHHOM CTPYKTYPhJ. Pa3JIH'-laJOT 4 ypoBHH CTPYKTYPHOM opratt113aUH11 6eJIKOB, Ha3brnaeMhix nepBH'-IHOM, BTOpH'-IHOtt, TpeTHllHOtt II qernepTH'-1- HOH CTpYJ<TYpaMII (pIIc. Cymecrny10T 06m11e rrpaBHJia, no KOTOPhIM H.LI,er ¢opMHpoBaHHe rrpocTpattcTBeHHhIX CTPYKTYP 6eJIKOB.

PMC. 1-3. 3Tanbl lj>opMMpOBSHMSI KOH!j>OpM84MM 6enKOB.

1 - nepBM4HaR CTPYKTYPa; 2 - BTOPM4Hal'!CTPYKTYpa; 3 - TpeTL'14Hal'!CTPYKT)'pa;4 - 40TBepTM4Hal'ICTpyKT)'pa.

19

«DepBH'l-

A. IIEPBifqHAfI CTPYKTYPA

AMHHOKHCJIOTHhie OCTaTKH B nenTH,!I.HOH uenH 6eJIKOB qepe,II.yIOTCH He CJiyl.JaHHhIM o6pa30M, a pacnoJIO)J(etthI B onpe,II.eJiettttoM nopMKe. Jlttttetl-

HYIO TIOCJie,II.OBaTeJihHOCTb aMHHOKHCJIOTHbIX OCTaTKOB B IlOJIHTieIITH,!I.HOH uenH Ha3bIBaIOT

uas CTPYKTYPa 6eJIKa».

TiepswrnaH CTPYKTYPa Ka)J(,!I.Oro HH,II.HBH,II.yaJibHOro 6eJIKa 3aKO,II.HpOBaHa B yqacTKe ,11,HK, Ha3bIBaeMOM rettoM. B rrpouecce CHHTe3a 6eJIKa HttcpopMaUHH, HaXOMlUaHCH B reHe, CHa<JaJia nepenHChIBaeTCH Ha MPHK, a 3aTeM, HCIIOJib3YH MPHK B Ka<Jecrne MaTpHUbI, Ha pH60COMe IIPOHCXO,II.HT c6opKa nep- Bl1'-!HOHCTpYKTYPhl 6eJIKa (CM. pa3,II.eJI 4).

Ka)J(,II.hIH H3 50 000 HH,II.HBHAYaJihHbIX 6eJIKOB oprattH3Ma '-!eJIOBeKaHMeeT yttHKaJihHYIO AJI51 aaHHOro 6eJIKa rrepBH'IHYIOCTPYKTYPY· Bee MOJieKyJibI ,II.aHHOfO HHAHBH.llYaJibHOfO 6eJIKa HMeIOT O,!I.HHaKOBOe qepeaoBaHHe aMHHOKHCJIOTHhIX OCTaTKOB B 6eJIKe, 'ITO B nepBYfO O'IepeAb OTJIH<JaeT aaHHblll HH)ll1BH,II.yaJibHbIH 6eJIOK OT mo6oro apyroro.

Ii. METO.IJ.hI IIJyqEHIIH nEPBIIqHoiI

CTPYKTYPhl BF.JIKA

lfayqeHHe nepBHl.JHOtt CTPYKTYPhI 6eJIKOB 11MeeT Ba)J(HOe o6me6HOJIOfH'leCKOe H MeAHUHHCKOe 3Ha'leHHe.lfayqaH nopMOK qepeaosaHHH aMHHOKHCJIOTHhIX OCTaTKOB B HHAHBHAYaJibHhlX 6eJIKax H COTIOCTaBmrn 3TH 3HaHH51 c oco6eHHOCTHMH npoCTpaHCTBeHHOro paCIIOJIO)J(eHH51 MOJieeyJihl, MO)J(- HO Bbl51BHTh o6mtte cpyHAaMeHTaJihHhie 3aKOHOMepHOCTH cpopMHPOBaHH51 npocTpaHCTBeHHOH CTPYKTYPbI 6eJIKOB.

KpoMe Toro, MHor11e reHeTHl.JecKHe 6oJie3HH - pe3yJibTaT HapyweHH.51 B aMHHOKHCJIOTHOH TIOCJie-

.llOBaTeJihHOCTH 6eJIKOB. HHcpopMaUHH o nepBHl.J- Hoii CTPYKTYPe HOPMaJihHoro 11 MyTaHTHoro 6eJIKa MO)J(eT 6hITb noJie3Ha AJIH )lH3fHOCTHKH H nportto311POBamrn pa3BHTHH 3a6oJieBaHl151.

YcrnHoBJiemi:e neps11'-!Hoi1 CTPYKTYPhI 6eJIKOB BKJIIO'IaeT2 OCHOBHbIX 3Tana:

• onpe)leJiemte aMHHOKHCJIOTHOfO COCTaBa ll3YqaeMoro 6eJIKa;

• orrpe,II.eJieHHe aMHHOKl1CJIOTHOH TIOCJieAOBaTeJibHOCTH B 6eJIKe.

]. OnpeiJe11enue QMUHOKUCJlOmROW COCma6a OettKa

nepBbIH 3TaTI B onpeaeJieHl111 nepBH'IHOHCTPYK-

TYPbl 6eJIKOB 3aKJIIOl.JaeTCH B Ka'IeCTBeHttoti11 Ko-

JIHl.JecrneHHOH oueHKe aMHHOKHCJIOTHOfO COCTaBa JJaHHoro HHAHBHAYaJihHoro 6eJIKa. Heo6xoawMo TIOMHHTh, l.JTO AJIH 11CCJieAOBClHHH H~HO HMeTb onpeneJieHHOe KOJIH'IeCTBO '-IHCTOfO 6eJIKa, 6e3 np11Mecetl ,II.pyrwx 6eJIKOB HJIH nenTHLJ.OB.

KuCJIOTHbIH rH,!J,pOJIH3 6eJIKa

Ll,JIH onpeneJieHH51 aMHHOKHCJIOTHOfO COCTaBa

Heo6XO,II.HMO rrpoBeCTH pa3pymeHHe Bcex nenTH)J,- HbIX CB513ett B 6eJIKe. AttaJIH3HpyeMhIH 6eJIOK mnPOJil13YIOT B 6 MOJI/JI HCI rrp11 TeMrreparype OKOJIO

110 °C B Tel.JeHwe 24 'I.B pe3yJibTaTe TaKOH o6pa6oTKH pa3pywaIOTCH rrenTH,II.Hbie CB51311 B 6eJIKe, a B fl1,!I.pOJIH3aTe rrpHCyTCTBYfOT TOJihKO CB060,II.Hb!e aM11HOKHCJIOThI. KpoMe Toro, rnyTaMHH 11 acrraparwH rH,II.pOJil13YIOTCH no fJiyTClMHHOBOH H acnaparHHOBOH KHCJIOT (T.e. pa3pbrnaeTCH aMH,II.HaH CBH3h B PM11KaJie H OT HHX onuerrJI51eTCH aM11Horpyrrrra).

Pa:meJieuue aMHHOKHCJIOT c noMO~IO

HOH006MeHHOH xpoMaTorpacJ>uu

CMecb aMHHOKHCJIOT, rroJiy'-!eHHhIXKHCJIOTHhIM mnpoJIH30M 6eJIKOB, pa3)J,eJIHJOT B KOJIOHKe c KaTHOH006MeHHOH CMOJIOH. TaKaH CHHTeTWieCKaH CMOJia conep)J(HT rrpO'-IHOCB513aHHhie c Hett OTpHuaTeJihHO 3ap5I)J(eHHhie rpyrrnhI (ttarrpHMep, ocrnTKH CYJibcpOHOBOH KHCJIOThl .. SQ3- ), K KOTOpbIM rrp11coenwHeHhI HOHbl Na+ (p11c. 1-4).

B KaTwottoo6MeHHHK BHOCJIT cMeCh aMHHOKHCJIOT B KHCJIOH cpene (pH 3,0), rne aMHHOKHCJIOTbl B OCHOBHOM rrpe,II.CTaBJIHIOT KaTHOHhl, T.e. HeCyT rroJIO)J(HTeJibHhlll 3apH,!I.. TioJIO)J(l1TeJibHO 3ap5I)J(etttthie aMHHOKHCJIOTbI rrpwcoenwHHIOTCH K 0Tp11uaTeJibHO 3apH)J(eHHhIM qacTHUaM CMOJibl. 1IeM 6oJihwe CYMMapHhiii 3ap51LI, aMHHOKHCJIOThl, TeM npOl.JHee ee CBH3b co cMoJiott. TaK, aMHHOKHCJIOThI JIH3Htt, aprnHHH H fHCTHLl,HH HaH60Jiee npol.JHO CBH3bIBaIOTCH c KaTHOH006MeHHHKOM, a acrrapamHOBaH H rnyTaMHHOBaH KHCJIOThI - Ha1160Jiee CJia6o.

Bb1cso6o)J(,II.eHwe aMHHOKHcrroT H3 KOJIOHKH ocymecTBJIHIOT BblMhIBaHMeM (::WIOHpOBaHHeM) HX 6ycpepHhIM paCTBOpOM c yBeJIHl.JHBaIOmetiCH HOHHOH cHJioti (T.e. c ysenwqettHeM Koliu,eHTpau,1111 NaCl) 11 pH. Tip11 ysenwqemrn pH aMHHOKHCJIOThI Tep51IOT npOTOH, B pe3yJihTaTe YMeHbmaeTCH HX ITOJIO)J(HTeJihHhIH 3apH,!I., a CJie)J,OBaTeJihHO H rrpoq - HOCTh CB513H c OTPHUaTeJihHO 3ap5I)J(eHHhIMH qacTHUaMH CMOJibl.

Ka)J(,!I.aH aMHHOKHCJIOTa BhIXO,II.HT H3 KOJIOHKH npw onpeneJieHHOM 3Ha'-lettw11pH n 11ottHoil CHJihI. Co-

20

.....

 

 

 

 

 

 

6y¢ep

 

 

 

 

 

t-CMeCb

 

 

 

 

 

 

 

0

 

 

8MlllHOKlllCilOT

 

 

 

 

 

 

 

 

0

 

0

 

 

 

 

 

0

t- KarnoHoo6MeHHlllK

 

 

 

 

 

 

 

 

0

 

 

 

 

0

 

0

 

 

 

 

0

°

0

 

 

 

 

0

 

 

 

 

 

0

0

0

 

 

 

0

 

 

0

 

 

 

 

 

 

 

 

l1Il:t;:t;IID

C6op

 

 

LJ

/

:mtaara

D

-11-11-n-n- KonneKTop

,1:1m1 c6opa lliillll cppaKUllli;j

A

+NH3+- CH - COOH

I

R (3ap5I,ll 0 lllJllll +)

6

PMc. 1·4. Pa3AE!JleH1te aMMHOK1tcnoT c noMO~blO MOHoo6MeHHOM xpoMaTorpacj>MM. A. XpoMaTorpacpw-1ecKafl KOflOHKa, HanonHeHHafl K8TlllOH006MeHHOiil CMonoiil. 6. 8Tanbl pa:meneHlllSI aMlllHOKlllCflOT: 1 - np111coei:1111HeH111e aMlllHOKlllCflOT K '"18CTlllU8MCMOflbl; 2 - BblCB060>K- AeH111e aM111HoK111cnoT np111 onpei:ieniiHHOM 3Ha'"leH111111pH 111 KOHUeHrpau111111 NaCl.

6ttpaH c HlDKHero KOHUa KOJIOHKH paCTBOP (3JIIOaT)

B B.H,LJ:e He60JihlllMX IIOpUMM, MO)!(HO IIOJIY'IMTb<}>paK-

UMM, COLl,epxall.IJ1e OTJJ,eJibHhie aMHHOKHCJIOThl.

KoJIJ111ecreeHHb1if aHaJIH3 nOJiy11eHHbIX <l>PaKIUtii

Konw-recTBO KIDK,!1,0H 113 aMMHOKHCJIOT B JJ,aHHOM 6eJIKe onpeJJ,eJUIIOT, ttarpeBa.51 OTJJ,eJihtthie cl>paKUMH aMMHOKMCJIOT c HMHrttJ],pMHOM, o6pa3yIOIUJ1M coe.u11HeHMe Kpactto-<}>110neT0Boro UBern. HttTettcttBHocTh oKpacKH B rrpo6e rrponoputt:oHaJibHa Kon11qecTBy

Haxomimei1CH B Hett aMl1HOKl1CJTOTbI, Il03TOMY no cneKTPOcPOTOMeTp11qecKOMY H3MepeHMIO CBeTa, IIOfJIOllleHHOfO Hl1Hf.H,D:pl1HOBhIMl1 I1p0113BOLl,HbIMl1, MO)l(HO onpe.ueJTMTb co.uep)f(aHife KIDK,!1,0H aMHHOKHCJIOThl B r.H,D:pOJTH3aTe JJ,attttoro 6em<a.

B HaCTOHIUee BpeMH rrpouecc paJaenemrn tt KOJIWiecrneHHoro orrpe.uenett115I aMMHOKMCJIOT B r11npon113aTe 6eJTKa ITOJTHOCTb!O aBTOMaHJ3HpOBaH l1 ocymecrn1rneTcH B cneu11aJibHOM npu6ope - al\1HHOKHCJIOTHOM aHaJTH3aTope.

21

2. Onpeoe.!lenue OMunoKucAomnoii

nOCJ1eoo«ame.!lbHocrnu 6 Oe.!IKe

Onpe,lJ,e.JieHHe N-Konu:eeou aM11HOKHCJIOThI

e oeJIKe u nocJie,lJ,oeaTeJihHOCTll aMHHOKHCJIOT e oJI11roneDTH,lJ,ax

<I>emu:nnorn:ou;i-mttaT (<l>HTU) - peareHT, HCrrOJih3yeMhIH MH orrpe,lJ,eJTeHHH N-Kottuesotl aMHHOKHCJTOThI B rrerrTH,lJ,e. Ott cnoco6ett pearnposaTb c a-aMHHorpynnotl H a-Kap6oKCHJThHOH rpyrrnoil cBo6o,lJ,HhIX aMHHOKHCJTOT, a TaK>Ke c N-KoHuesort aMHHOKHCJIOTOH B nerrTH,lJ,ax (CM. CXeMY HH)!(e).

B pe1yJThTaTe B3aHMO,lJ,ei1cTBHH c N-KOHUeBoil aMHHOKHCJTOTOH ITOJTHTieITTH,lJ,a 06pa1yeTCH <PeHHJTTHOfH,lJ,aHTHOHOBOe rrpOH3BO,lJ,HOe, B KOTOPOM ,lJ,eCTa6HJTH3HpOBaHa rrerrTH,lJ,HaH CBH3b Me)!()ly CX-Kap- 60KCHJibHOH rpynrroit N-Kottuesoil aMHHOKHCJIOThr H a-aMHHorpynrrotl BTopotl aMHHOKHCJIOThI B rrerr - TH,lJ,e. 3rn CBH3b H36ttpaTeJTbHO rH,lJ,pOJIH3yeTCH 6e3 TIOBpe)!()leHIDI ,l1,pymx rreITTH,lJ,HbIX CBH3ei1.

+NH-CH- CO ... NH-CH- COOH

I

Rn

ITocne peaKUHH BhI,D;eJIHJOT KOMTIJTeKc <I>HTU-

AKv 11,11,eHTHqmuttpyIOT ero xpoMaTorpaqrnqecKHMH MeTO)JaMH. <I>HTU MO)!(HO HCITOJlb30BaTb BHOBb c YKOpoqeHHhIM rrenTH,lJ,OM, nonyqeHHhIM B npe- ,lJ,hI,D;YIUeM 11,HKJTe, MH orrpe,lJ,eJieHHH CJie,lJ,yJOill,eti aMHHOKHCJIOThI. 3TOT rrpouecc cTyrrettqaToro pacrueTIJieHHH rrenTH,lJ,a c N-Kottua 6hm aBTOMaTH3H - poBaH H peaJTH30BaH B rrptt6ope - ceKBeHaTope, c TIOMOIUhJO KOToporo MO)!(HO onpe,lJ,eJIHTh TTOCJie,lJ,O- BaTeJibHOCTb aMHHOKHCJIOTHhIX OCTaTKOB B OJTHfOneTITH,lJ,aX, COCTOHIUHX H3 10-20 aMHHOKHCJIOT.

Mttortte nonHnerrTH,lJ,hI HMeJOT rrepBHY:HYJO CTPYKTYPY, cocTOHIJIYIO 6oJTee qeM H3 100 aMHHO-

KHCJTOT. TaK KaK c noMOIIJ,hJO ceKBeHaTopos HaH6onee rrpo)lyKTHBHO onpe,l1,eJ15IlOT aMJIHOKHCJIOTHYJO ITOCJTe,lJ,OBaTeJThHOCTb JIHillb He60JTbllIHX rrerrTH,lJ,OB, MOJieKYJThI noJIHrrenTH,lJ,a pacu~errAAJOT rro cneuHqrnqecKHM MeCTaM Ha ¢parMeHTbI.

l1crrOJib3YH HeCKOJlbKO pa3HbIX pacmerrJTHIOill,HX areHTOB (HMH MOryT 6hITb <f>epMeHTbI HJTH XHMHqecKHe seu~ecTBa) B pa3HhIX npo6ax oqwruettttoro ITOJTHITeTITH,lJ,a, MO)!(HO TIOJiyqHTb qaCTHqHo nepeKpbIBaJOilIHe DPYf ,lJ,pyra <f>parMeHThI c ycrnttoBJieHHOil aMHHOKHCJIOTHOH ITOCJTell,OBaTeJThHOCTbJO. c

HX TIOMOillhlO MO)!(HO BOCC03,lJ,aTb rrpaBHJlbHhlH nopH,lJ,OK <f>parMeHTOB H TIOJiy'IHTbTIOJTHYIO TIOCJie11,0BaTeJThHOCTb aMHHOKHCJTOT B TIOJTHITeTITH,lJ,HOH uen11.

<I>epMeuTaTHeuoe pacmeDJiemte noJIHIIeDTH,lJ,a no cneuucl>WiecKHM Y'laCTKaM

,Ll,.rrH crreutt<f>ttqecKoro pacIUenJTeHHH rrenTH,lJ,HhIX CB513eH B 6eJTKe MO)((HO HCITO.Jih30BaTh HeCKOJlhKO pa3HhIX ¢epMettTOB. Han6onee llIMpOKO 11crroJTh1y10T ¢epMeHTaTHBHblH fH,l1,pOJTH3 ITOJIHTieITTH,lJ,a rrpoTeOJTHTHqecKHM <f>epMeHTOM - TPHITCHHOM, KOTOpbIH OTHOCHT K rpyrrne ITHilleBaptt:TeJibHbIX <f>epMeHTOB ( ero Bhipa6aThrnaeT rrO)l)!(eJTyJJ:oqttaH )!(eJTe3a). <I>epMettT o6n3,lJ,aeT BhICoKoil cneutt<f>HqHOCThJO ,lJ,eticTBHH. Ott pacu~errJTHeT rrerrTH,lJ,Hhie CB513H, B o6pa30BaHHH KOTOpbIX yqacrnyeT Kap6oKCHJlbHa5l rpynrra OCTaTKOB JTH3HHa HJIH apmHHHa.

-NH-CH-CO-NH-CH-CO-

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J1Lll3

 

 

111n111Apr

HCXOll,51 H3 ycTaHOBJieHHOfO KOJTH'leCTBaOCTaTKOB JlH3MHa H aprMHMHa, MO)!(HO npe,l1,CKa3aTh KOJIMqecTBO ITOJTyY:aeMhIX npH fH,l1,pOJTH3e TPHTICHHOM <f>parMeHTOB. TaK, ecJTH B rroJTttrrerrTH,lJ,HOH uerrH

6 OCTaTKOB aprHHHHa H JTH311Ha, TO rrpH pacmerr-

0-N=C=S + H2N-CH-CO-NH-CH-CO ... -NH-CH-COCH

 

I

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R1

R2

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1

nenrn.1:1

 

CxeMa

22

1-5).

nett1111 Tp11nc11HOM MO)l(HO nonyqttTh 7 cf:>parMeHTOB.

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XnMH'l.ecKoepacmern1euue no;mnennt,11,a no cneuucl>HqecKHM yqacncaM

B HeKoTophIX cny1rn5lx npennoYTHTeneH He cf:>epMeHTaTHBHhitt, a x11M1111ecKHi1 mnpon11J. TaK, pearettT 6pOMUHaH paClllef1JI5leT TOJlbKO rrenTH)lHhie CB5l3l1, B KOTOpblX Kap60KCl1JlbHaH rpynna npHtta)lJie)f(l1T OCTaTKY MeTHOHHHa. 3HaH KOJllflieCTBO OCTaTKOB MeTMOHHHa B ITOJIHrrenTHJIHOH uerrH, JTerKO ycTaHOBHTb KOJTH'leCTBOnony11aeMbIX cf:>parMeHTOB. 11,anee JIJIH Ka)l()loro cf:>parMeHTa B ceKBettaTope TaIOKe ycrnHaBJTHBalOT aMl1HOKHCJIOTHYJO nocJTeJlOBaTeJibHOCTb.

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11,Jm ycrreurnoro ycrnHoBJieHHH nocne)lOBaTeJibHOCTH nonyi1ettHhIX cf:>parMeHTOB nommenn1na He06xon.11Mo nony11.11Th nenTHJlhI c nepeKphIBalOWHMHCH aMHHOKl1CJIOTHb!Ml1 flOCJie)lOBaTeJihHOCTHMH. 3TO JJ,OCTHfalOT o6pa60TKOH OTJJ,eJThHbIX npo6 naHHOfO noJIMnenTH)la pa3HhIMl1 peareHTaMtt, paclllenJIHIOliIMMH 6enoK B pa3HhIX Mecrnx. Heo6xo)lttMO npoBeCTH CTOJlbKO paclllenJTeHHH, 4T06br noJTyqHTb tta6op nerrTH)lOB, 06ecrre1nrna10wttx nepeKphrnaH11e scex y 1rncTKOB, tteo6xon11MhIX JJ,JIJJ onpeJJ,eJTeHttH TIOCJle)lOBaTeJibHOCTH HCX0).1,HOfO TIOJTHTieITTH)la.

Ana-r11c-Apr-npo-MeT

YcTaHosneHMe nepBM'-IHOMCTPYKTYPbl 6enKa c noMOIJ..lblO nepeKpb1Ba1011..1Mxcsi nenTM1'HbtX<t>parMeHTOB.

B. Kott<DOPMAU,m1 liE.JIKOB

JlttHeHHbie ITOJIHnerrTH)lHhie uem1 11HlJ:l1BH)lYaJTbHblX 6eJIKOB Ja C'leT B3aHMO)leHCTBHH ¢YHKIIHOHaJTbHblX rpyrrrr aMHHOKHCJIOT rrptto6peTalOT onpeneJieHHYlO npOCTpaHCTBeHHYIO TpexMepHylO CTPYKTypy, tta3bIBaeMyIO «Kou<t>opMau,u.H.». Bee MO-

JTeKyJihl 11H,lll1Bl1).1,yaJThHbIX 6eJIKOB (T.e. ttMeIOliIHX OJ1l1HaKOBYJO nepB1111tty10 CTPYKTYPY) 06pa3y10T B pacrnope OJ1HHaKOBYJO Kottcf:>opMau11IO. Cne)losaTeJThHO, BCH HH¢opMaUH5J, He06XO)ll1Ma5l J1,JlH cf:>opMHpOBaHHH npocTpattcTBeHHhIX cTpyKTyp, ttaxoJ1HTCH B rreps1111ttoi1 crpyKType 6eJIKOB.

B 6eJIKax pa3JTHlJaIOT 2 OCHOBHblX THna KOHcpopMaUHH ITOJIHITeITTH)lHbIX uerreJ:r: BTOPHllHYIO H TpeTWIHYIO CTPYKTYPhI.

1. BmopU'IHOJl cmpy«mypa 6e.riKo6

BTOpH'luanCTJ>YJITYPa 6eJIKoe - rrpocrpaHcTBeHHaH crpyKzypa, 06pa3y101UaHCH B pe3yJihTaTe B3a11MOJ1ettcrn11« Me)l()ly cf:>yttKI1110HaJTbHhIMl1 rpynnaM11, BX0).1,HIUHMH B COCTaB neIITl1).1,HOfO OCTOBa. CTpH 3TOM rrenTHJ1Hh1e uentt Moryr nptto6peTaTh perym1ptth1e crpyKTYPhI JIByx Tttrros: a-crr11panb 11 ~-cTpyKTypa.

a-CnupaJib

B )laHHOM Tttrre CTPYKTYPhI rrerrTHJ1Hhitt ocTOB JaKpyirnsaeTcJI B BH)le cnnparrtt 3a cqfa 06pa3osaHHJI BOJIOPOJ1HblX CB513eH Me)[():(y aTOMaM11 KHCJIOpo)la Kap60Hl1JibHbJX rpynn H aTO_MaMH a30Ta aMHHOrpyrrn' BX0).1,HliIHX B COCTaB rrerrTH)lHbIX rpyrrrr qepe3 4 aMHHOKttCJTOTHhIX ocTaTKa. BonoponHhie CB513H optteHTttpoBaHhl B)lOJlb OCH crrttpaJTH (p11c.

Ha O)ltttt BHTOK a-crrttpam1 npttxo)lttTCH 3,6 aMHHOKHCJIOTHblX OCTaTKa.

B 06pa3osattm1 BO)lOpO)lHhIX CBH3ett yqacrnyioT rrpaKTl14eCKl1 BCe aTOMbl KHCJIOpO)la 11 BOJlOPOUa nenTH)lHblX rpyrrrr. B pe3yJibTaTe a-crrnparrh «cnrr11BaeTC5J» MHO)l(eCTBOM BOJ10POJ1Hh!X CBH3e:ti. HecMOTp5l Ha TO, lJTO )laHHhie CB513H OTHOCHT K PaJPHIIY CJia6bIX, 11X KOJTH'IeCTBO 06ecne1111BaeT MaKCHMaJTbHO BOJMO)KJ-!YIO crn6HJihHOCTh a-cn11pantt. TaK KaK Bee fH)lpocpHJihHbie rpynTihI nerrTl1,l1,HOfO OCTOBa 06bl4HO yqacTBYJOT B o6pa30BaHl111 BOJ10POI1HhIX CBHJe:ti, rl1).1,po¢HJibHOCTb (T.e. cnoco6HOCTh o6pa30BhIBaTh BO)lOpO)lHbie CBH3l1 c BOJlOH) a-crmpanett yMeHhWaeTCH, a l1X fH)lpocf:>o6HOCTb yBeJIH'111BaeTC5l.

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Paa11KaJThI aM11HoK11cnoT ttaxOMTCH tta ttap~Hott

CTOPOHe a-cnttpaJIH 11 HanpaBJieHbl OT rrerrTH)lHOfO OCTOBa B CTOpOHbl. 0HH He yqaCTBYJOT B o6pa30Ba-

23

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